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Article(id=1200147842271637877, tenantId=1146029695717560320, journalId=1190317699101192196, issueId=1200147837586604797, articleNumber=1001-2494(2024)10-0887-09, orderNo=null, doi=10.11669/cpj.2024.10.005, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1680019200000, receivedDateStr=2023-03-29, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1764067143266, onlineDateStr=2025-11-25, pubDate=1716307200000, pubDateStr=2024-05-22, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1764067143266, onlineIssueDateStr=2025-11-25, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1764067143266, creator=13701087609, updateTime=1764067143266, updator=13701087609, issue=Issue{id=1200147837586604797, tenantId=1146029695717560320, journalId=1190317699101192196, year='2024', volume='59', issue='10', pageStart='857', pageEnd='950', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1764067142149, creator=13701087609, updateTime=1764067345188, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1200148689244225889, tenantId=1146029695717560320, journalId=1190317699101192196, issueId=1200147837586604797, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1200148689244225890, tenantId=1146029695717560320, journalId=1190317699101192196, issueId=1200147837586604797, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=887, endPage=895, ext={EN=ArticleExt(id=1200147842632348027, articleId=1200147842271637877, tenantId=1146029695717560320, journalId=1190317699101192196, language=EN, title=Chemical Identification of TCM Sargassum Based on Characteristic Fatty Acids, columnId=null, journalTitle=Chinese Pharmaceutical Journal, columnName=null, runingTitle=null, highlight=null, articleAbstract=

OBJECTIVE To study the identification of Sargassum fusiforme and other sargassos (algae from the genus Sargassum) by targeted lipidomics combined with chemotaxonomy methods. METHODS Twenty-seven batches of S. fusiforme were collected and the GC-MS fingerprint of fatty acids (FAs) was established. The principal component analysis and orthogonal partial least squares discriminant analysis were used to compare S. fusiforme with other 8 species, total 22 batches of sargassos. Based on the differential FAs between S. fusiforme and other sargassos, combined with the analysis of FA biosynthesis pathway, the Q-marker was screened. RESULTS A total of 29 common FAs were identified by the fingerprint, and the similarity was greater than 0.96. The chemical pattern recognition method can well distinguish S. fusiforme from other 8 species, and the discrimination model established by this method can accurately verify 15 batches of commercial slices. Based on the analysis of differential FAs and FA biosynthesis pathway, monounsaturated fatty acid was found to play an important role in chemical classification. The ratio C20∶1 n-9/C16∶1 n-7 was proposed to be used as a candidate Q-marker for the identification of S. fusiforme and 14 batches of commercial slices can be accurately verified. CONCLUSION This study establishes the identification model of S. fusiforme and other confused sargassos, finds the Q-marker of FAs, and provides a reference for the establishment of the identification method of TCM SARGASSUM.

, correspAuthors=Lidong LIN, Hongbing LIU, authorNote=null, correspAuthorsNote=null, copyrightStatement=null, copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=null, magXml=null, pdfUrl=null, pdf=null, pdfFileSize=null, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=null, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=null, mapNumber=null, authorCompany=null, fund=null, authors=null, authorsList=Xicheng GENG, Shanpeng MA, Jiapeng CHEN, Lujing CHEN, Lidong LIN, Hongbing LIU), CN=ArticleExt(id=1200147844289098181, articleId=1200147842271637877, tenantId=1146029695717560320, journalId=1190317699101192196, language=CN, title=基于特征脂肪酸的中药海藻鉴别研究, columnId=1190352405612040510, journalTitle=中国药学杂志, columnName=论著, runingTitle=null, highlight=null, articleAbstract=

目的 利用植物脂质组学和化学分类学开展羊栖菜与其同属易混马尾藻的鉴别研究。方法 收集27批羊栖菜,靶向建立了脂肪酸气相色谱-质谱(GC-MS)指纹图谱;运用主成分分析、正交偏最小二乘判别分析等化学模式识别方法,比较了羊栖菜与同属其他8种22批马尾藻,建立了海藻饮片判别模型;基于羊栖菜与其他马尾藻的差异脂肪酸,结合脂肪酸生合成途径分析,筛选质量标志物。结果 指纹图谱共标定29个共有脂肪酸,相似度均大于0.96。化学模式识别法能够很好区分羊栖菜与同属其他8种22批马尾藻,以此建立的判别模型能够准确验证15批市售海藻饮片。结合差异脂肪酸与脂肪酸生合成途径分析,发现单不饱和脂肪酸在化学分类中的重要作用,提出比值C20∶1 n-9/C16∶1 n-7可作为羊栖菜鉴别的候选质量标志物,14批市售海藻饮片可被准确验证。结论 本研究建立了羊栖菜与同属易混马尾藻的判别模型,发现了脂肪酸质量标志物,为中药海藻真伪鉴别方法的制定提供参考。

, correspAuthors=林立东, 刘红兵, authorNote=null, correspAuthorsNote=
*林立东,男,博士,高级工程师 研究方向:基础生物学 Tel: (0577)59386761;
刘红兵,女,博士,教授,博士生导师 研究方向:海洋中药药效物质及质量评价研究 Tel: (0532)82031823
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耿玺承,女,硕士研究生 研究方向:海洋中药

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耿玺承,女,硕士研究生 研究方向:海洋中药

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耿玺承,女,硕士研究生 研究方向:海洋中药

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4 NMPA Key Laboratory for Quality Research and Evaluation of Marine Traditional Chinese Medicine, Qingdao 266000, China, bio=null, bioImg=null, bioContent=null, aboutCorrespAuthor=null), CN=AuthorExt(id=1200147847741010499, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1200147842271637877, authorId=1200147847384494649, language=CN, stringName=刘红兵, firstName=null, middleName=null, lastName=null, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=1, 2, 4, *, address=1 中国海洋大学, 海洋药物教育部重点实验室, 山东 青岛 266003
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J Fish China(水产学报), 2020, 44(4):581-595., articleTitle=Supplementary study on reproductive biology and life cycle of Sargassum fusiforme(Phaeophyceae), refAbstract=null)], funds=[Fund(id=1200147851742376623, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1200147842271637877, awardId=81973433, language=CN, fundingSource=国家自然科学基金项目资助(81973433), fundOrder=null, country=null), Fund(id=1200147851901760178, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1200147842271637877, awardId=HYZY-ZZKT-2022002, language=CN, fundingSource=国家药品监督管理局海洋中药质量研究与评价重点实验室课题资助(HYZY-ZZKT-2022002), fundOrder=null, country=null), Fund(id=1200147851973063348, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1200147842271637877, awardId=2022QNLM030003-2, language=CN, fundingSource=青岛海洋科技中心专项资助(2022QNLM030003-2), fundOrder=null, country=null)], companyList=[AuthorCompany(id=1200147844540756436, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1200147842271637877, xref=1, ext=[AuthorCompanyExt(id=1200147844549145044, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1200147842271637877, companyId=1200147844540756436, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=1 Key Laboratory of Marine Drugs, Ministry of Education, Ocean University of China, Qingdao 266003, China), AuthorCompanyExt(id=1200147844553339349, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1200147842271637877, companyId=1200147844540756436, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=1 中国海洋大学, 海洋药物教育部重点实验室, 山东 青岛 266003)]), AuthorCompany(id=1200147844633031131, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1200147842271637877, xref=2, ext=[AuthorCompanyExt(id=1200147844637225436, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1200147842271637877, companyId=1200147844633031131, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=2 Laboratory for Marine Drugs and Bioproducts, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266237, China), AuthorCompanyExt(id=1200147844645614045, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1200147842271637877, companyId=1200147844633031131, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=2 海洋药物与生物制品功能实验室, 青岛海洋科学与技术试点国家实验室, 山东 青岛 266237)]), AuthorCompany(id=1200147844716917222, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1200147842271637877, xref=3, ext=[AuthorCompanyExt(id=1200147844721111524, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1200147842271637877, companyId=1200147844716917222, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=3 Marine and Fishery Development Research Center of Dongtou District, Wenzhou 325700, China), AuthorCompanyExt(id=1200147844729500133, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1200147842271637877, companyId=1200147844716917222, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=3 温州市洞头区海洋与渔业发展研究中心, 浙江 温州 325700)]), AuthorCompany(id=1200147844817580520, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1200147842271637877, xref=4, ext=[AuthorCompanyExt(id=1200147844821774826, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1200147842271637877, companyId=1200147844817580520, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=4 NMPA Key Laboratory for Quality Research and Evaluation of Marine Traditional Chinese Medicine, Qingdao 266000, China), AuthorCompanyExt(id=1200147844825969132, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1200147842271637877, companyId=1200147844817580520, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=4 国家药品监督管理局海洋中药质量研究与评价重点实验室, 山东 青岛 266000)])], figs=[ArticleFig(id=1200147849255154286, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1200147842271637877, language=EN, label=Fig.1, caption=GC-MS analysis on fatty acids of S. fusiforme

A-fatty acid fingerprint of S. fusiforme; B-chromatogram of fatty acid methyl ester reference substance peak; Peak 1-29-conmmon fatty acid peaks of 27 batches of S.fusiforme; S1-S27-S.fusiform SF01-SF27; R-reference fingerprint.

, figureFileSmall=99Ua+rnfUn9YFdsBrHBEBA==, figureFileBig=iwIzm+WVuD04RXx0noIpew==, tableContent=null), ArticleFig(id=1200147849339040370, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1200147842271637877, language=CN, label=图1, caption=羊栖菜脂肪酸气相色谱-质谱(GC-MS)图

A-羊栖菜脂肪酸指纹图谱;B-脂肪酸甲酯对照品图谱峰;峰1~29-27批羊栖菜共有脂肪酸峰;S1~S27-羊栖菜SF01~SF27;R-对照指纹图谱。

, figureFileSmall=99Ua+rnfUn9YFdsBrHBEBA==, figureFileBig=iwIzm+WVuD04RXx0noIpew==, tableContent=null), ArticleFig(id=1200147849599087221, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1200147842271637877, language=EN, label=Fig.2, caption=Principal components analysis score plot of S. fusiforme and other sargassos, figureFileSmall=OetV5/yxkLgz8B8oD5SfIg==, figureFileBig=ZLp2FR/TeMJN/ixYFuSxQQ==, tableContent=null), ArticleFig(id=1200147849729110647, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1200147842271637877, language=CN, label=图2, caption=羊栖菜及同属他种马尾藻脂肪酸的主成分分析(PCA)得分图, figureFileSmall=OetV5/yxkLgz8B8oD5SfIg==, figureFileBig=ZLp2FR/TeMJN/ixYFuSxQQ==, tableContent=null), ArticleFig(id=1200147849867522682, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1200147842271637877, language=EN, label=Fig.3, caption=Orthogonal partial least squares-discriminant analysis of S. fusiforme and other sargassos

A-permutation test; B-OPLS-DA score plot; C-S-plot; D-differential fatty acid contents; 1)P<0.05, 2)P<0.01, compared with S.fusiforme.

, figureFileSmall=nsh4eBmdBxv0PTaR/X/X1g==, figureFileBig=KANuAshMGlqbpqATV5OXTg==, tableContent=null), ArticleFig(id=1200147850056266366, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1200147842271637877, language=CN, label=图3, caption=羊栖菜及同属他种马尾藻脂肪酸的正交偏最小二乘判别分析(OPLS-DA)图

A-置换检验图;B-得分图;C-S-plot图;D-差异脂肪酸含量图;与羊栖菜相比,1)P<0.05, 2)P<0.01。

, figureFileSmall=nsh4eBmdBxv0PTaR/X/X1g==, figureFileBig=KANuAshMGlqbpqATV5OXTg==, tableContent=null), ArticleFig(id=1200147850207261316, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1200147842271637877, language=EN, label=Fig.4, caption=Main road map of fatty acid biosynthesis pathways, figureFileSmall=MWoWav3uih5+YvzHBOmGBQ==, figureFileBig=0poHkQ/qzVcr9dl+W5FYxQ==, tableContent=null), ArticleFig(id=1200147850328896135, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1200147842271637877, language=CN, label=图4, caption=脂肪酸的生物合成途径主要路线图, figureFileSmall=MWoWav3uih5+YvzHBOmGBQ==, figureFileBig=0poHkQ/qzVcr9dl+W5FYxQ==, tableContent=null), ArticleFig(id=1200147850442142346, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1200147842271637877, language=EN, label=Fig.5, caption=Monounsaturated fatty acid ratio of S. fusiforme(n=27) and other sargassos(n=22). $\bar{x}±s$

1)P<0.01, compared with S. fusiforme.

, figureFileSmall=+xYHa0sbn3VdK68dbbeYWw==, figureFileBig=NxsthUmZNIkogMSV7guk4w==, tableContent=null), ArticleFig(id=1200147850567971470, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1200147842271637877, language=CN, label=图5, caption=羊栖菜(n=27)与同属他种马尾藻(n=22)的单不饱和脂肪酸相关比值. $\bar{x}±s$

与羊栖菜相比,1)P<0.01。

, figureFileSmall=+xYHa0sbn3VdK68dbbeYWw==, figureFileBig=NxsthUmZNIkogMSV7guk4w==, tableContent=null), ArticleFig(id=1200147850660246162, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1200147842271637877, language=EN, label=Tab.1, caption=

Information of sample of sargassos and Sargassum slices

, figureFileSmall=null, figureFileBig=null, tableContent=
Sample Sample number Place of origin (in Chinese) Acquisition time Notes
Sargassum fusiforme SF01 Wenzhou,Zhejiang,China(中国浙江温州) 2021.01 Wild
Sargassum fusiforme SF02 Rongcheng,Shandong,China(中国山东荣成) 2021.01 Wild
Sargassum fusiforme SF03 Wenzhou,Zhejiang,China(中国浙江温州) 2021.01 Wild
Sargassum fusiforme SF04 Nan'ao,Guangdong,China(中国广东南澳) 2020.02 Cultured
Sargassum fusiforme SF05 Rongcheng,Shandong,China(中国山东荣成) 2020.02 Wild
Sargassum fusiforme SF06 Wenzhou,Zhejiang,China(中国浙江温州) 2020.02 Wild
Sargassum fusiforme SF07 Wenzhou,Zhejiang,China(中国浙江温州) 2020.02 Cultured strain-1
Sargassum fusiforme SF08 Wenzhou,Zhejiang,China(中国浙江温州) 2020.02 Cultured strain-2
Sargassum fusiforme SF09 Wenzhou,Zhejiang,China(中国浙江温州) 2020.02 Cultured strain-3
Sargassum fusiforme SF10 Wenzhou,Zhejiang,China(中国浙江温州) 2020.03 Cultured strain-1
Sargassum fusiforme SF11 Wenzhou,Zhejiang,China(中国浙江温州) 2020.03 Cultured strain-2
Sargassum fusiforme SF12 Wenzhou,Zhejiang,China(中国浙江温州) 2020.03 Cultured strain-3
Sargassum fusiforme SF13 Nan'ao,Guangdong,China(中国广东南澳) 2020.03 Wild
Sargassum fusiforme SF14 Rongcheng,Shandong,China(中国山东荣成) 2020.03 Cultured
Sargassum fusiforme SF15 Wenzhou,Zhejiang,China(中国浙江温州) 2020.03 Wild
Sargassum fusiforme SF16 Wenzhou,Zhejiang,China(中国浙江温州) 2020.04 Cultured strain-1
Sargassum fusiforme SF17 Wenzhou,Zhejiang,China(中国浙江温州) 2020.04 Cultured strain-2
Sargassum fusiforme SF18 Wenzhou,Zhejiang,China(中国浙江温州) 2020.04 Cultured strain-3
Sargassum fusiforme SF19 Wenzhou,Zhejiang,China(中国浙江温州) 2020.05 Cultured strain-1
Sargassum fusiforme SF20 Wenzhou,Zhejiang,China(中国浙江温州) 2020.05 Cultured strain-2
Sargassum fusiforme SF21 Wenzhou,Zhejiang,China(中国浙江温州) 2020.05 Cultured strain-3
Sargassum fusiforme SF22 Nan'ao,Guangdong,China(中国广东南澳) 2020.12 Cultured
Sargassum fusiforme SF23 Rongcheng,Shandong,China(中国山东荣成) 2020.12 Wild
Sargassum fusiforme SF24 Wenzhou,Zhejiang,China(中国浙江温州) 2020.12 Cultured
Sargassum fusiforme SF25 Nan'ao,Guangdong,China(中国广东南澳) 2020.12 Cultured
Sargassum fusiforme SF26 Wenzhou,Zhejiang,China(中国浙江温州) 2020.12 Wild
Sargassum fusiforme SF27 Wenzhou,Zhejiang,China(中国浙江温州) 2020.12 Cultured
S. pallidum SPA01 Yantai,Shandong,China(中国山东烟台) 2022.06 Wild
S. pallidum SPA02 Yantai,Shandong,China(中国山东烟台) 2022.06 Wild
S. thunbergii ST01 Wenzhou,Zhejiang,China(中国浙江温州) 2021.01 Wild
S. thunbergii ST02 Wenzhou,Zhejiang,China(中国浙江温州) 2020.01 Wild
S. thunbergii ST03 Wenzhou,Zhejiang,China(中国浙江温州) 2020.02 Wild
S. thunbergii ST04 Wenzhou,Zhejiang,China(中国浙江温州) 2020.03 Wild
S. thunbergii ST05 Wenzhou,Zhejiang,China(中国浙江温州) 2020.04 Wild
S. thunbergii ST06 Wenzhou,Zhejiang,China(中国浙江温州) 2020.06 Wild
S. thunbergii ST07 Wenzhou,Zhejiang,China(中国浙江温州) 2022.05 Wild
S. thunbergii ST08 Qingdao,Shandong,China(中国山东青岛) 2022.05 Wild
S. horneri SH01 Wenzhou,Zhejiang,China(中国浙江温州) 2020.02 Wild
S. horneri SH02 Wenzhou,Zhejiang,China(中国浙江温州) 2020.03 Wild
S. horneri SH03 Wenzhou,Zhejiang,China(中国浙江温州) 2020.04 Wild
S. horneri SH04 Wenzhou,Zhejiang,China(中国浙江温州) 2020.05 Wild
S. horneri SH05 Wenzhou,Zhejiang,China(中国浙江温州) 2020.06 Wild
S. horneri SH06 Wenzhou,Zhejiang,China(中国浙江温州) 2022.05 Wild
S. miyabei SM01 Yantai,Shandong,China(中国山东烟台) 2022.05 Wild
S. oligocystum SO01 Trat,Thailand(泰国达叻) 2019.02 Wild
S. oligocystum SO02 Trat,Thailand(泰国达叻) 2019.02 Wild
S. polycystum SP01 Chonburi,Thailand(泰国春武里) 2019.02 Wild
S. ilicifolium SI01 Ranong,Thailand(泰国拉农) 2019.01 Wild
S. binderi SB01 Chantaburi,Thailand(泰国尖竹汶) 2019.01 Wild
Sargassum slices YP01 Shandong,China(中国山东) 2018.05 S. fusiforme
Sargassum slices YP02 Shandong,China(中国山东) 2019.05 S. fusiforme
Sargassum slices YP03 Shandong,China(中国山东) 2021.05 S. fusiforme
Sargassum slices YP04 Guangxi,China(中国广西) 2019.05 S. fusiforme
Sargassum slices YP05 Shandong,China(中国山东) 2015.05 S. crassifolium
Sargassum slices YP06 Zhejiang,China(中国浙江) 2019.11 S. thunbergii
Sargassum slices YP07 Shandong,China(中国山东) 2019.12 S. crassifolium
Sargassum slices YP08 Fujian,China(中国福建) 2019.05 S. miyabei
Sargassum slices YP09 Fujian,China(中国福建) 2017.03 S. sp.
Sargassum slices YP10 Zhejiang,China(中国浙江) 2019.05 S. fusiforme
Sargassum slices YP11 Zhejiang,China(中国浙江) 2019.05 S. fusiforme
Sargassum slices YP12 Zhejiang,China(中国浙江) 2018.05 S. fusiforme
Sargassum slices YP13 Zhejiang,China(中国浙江) 2020.05 S. thunbergii
Sargassum slices YP14 Shandong,China(中国山东) 2015.05 S. horneri
Sargassum slices YP15 Shandong,China(中国山东) 2015.11 S. horneri
), ArticleFig(id=1200147850769298070, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1200147842271637877, language=CN, label=表1, caption=

马尾藻药材及中药海藻饮片样品信息表

, figureFileSmall=null, figureFileBig=null, tableContent=
Sample Sample number Place of origin (in Chinese) Acquisition time Notes
Sargassum fusiforme SF01 Wenzhou,Zhejiang,China(中国浙江温州) 2021.01 Wild
Sargassum fusiforme SF02 Rongcheng,Shandong,China(中国山东荣成) 2021.01 Wild
Sargassum fusiforme SF03 Wenzhou,Zhejiang,China(中国浙江温州) 2021.01 Wild
Sargassum fusiforme SF04 Nan'ao,Guangdong,China(中国广东南澳) 2020.02 Cultured
Sargassum fusiforme SF05 Rongcheng,Shandong,China(中国山东荣成) 2020.02 Wild
Sargassum fusiforme SF06 Wenzhou,Zhejiang,China(中国浙江温州) 2020.02 Wild
Sargassum fusiforme SF07 Wenzhou,Zhejiang,China(中国浙江温州) 2020.02 Cultured strain-1
Sargassum fusiforme SF08 Wenzhou,Zhejiang,China(中国浙江温州) 2020.02 Cultured strain-2
Sargassum fusiforme SF09 Wenzhou,Zhejiang,China(中国浙江温州) 2020.02 Cultured strain-3
Sargassum fusiforme SF10 Wenzhou,Zhejiang,China(中国浙江温州) 2020.03 Cultured strain-1
Sargassum fusiforme SF11 Wenzhou,Zhejiang,China(中国浙江温州) 2020.03 Cultured strain-2
Sargassum fusiforme SF12 Wenzhou,Zhejiang,China(中国浙江温州) 2020.03 Cultured strain-3
Sargassum fusiforme SF13 Nan'ao,Guangdong,China(中国广东南澳) 2020.03 Wild
Sargassum fusiforme SF14 Rongcheng,Shandong,China(中国山东荣成) 2020.03 Cultured
Sargassum fusiforme SF15 Wenzhou,Zhejiang,China(中国浙江温州) 2020.03 Wild
Sargassum fusiforme SF16 Wenzhou,Zhejiang,China(中国浙江温州) 2020.04 Cultured strain-1
Sargassum fusiforme SF17 Wenzhou,Zhejiang,China(中国浙江温州) 2020.04 Cultured strain-2
Sargassum fusiforme SF18 Wenzhou,Zhejiang,China(中国浙江温州) 2020.04 Cultured strain-3
Sargassum fusiforme SF19 Wenzhou,Zhejiang,China(中国浙江温州) 2020.05 Cultured strain-1
Sargassum fusiforme SF20 Wenzhou,Zhejiang,China(中国浙江温州) 2020.05 Cultured strain-2
Sargassum fusiforme SF21 Wenzhou,Zhejiang,China(中国浙江温州) 2020.05 Cultured strain-3
Sargassum fusiforme SF22 Nan'ao,Guangdong,China(中国广东南澳) 2020.12 Cultured
Sargassum fusiforme SF23 Rongcheng,Shandong,China(中国山东荣成) 2020.12 Wild
Sargassum fusiforme SF24 Wenzhou,Zhejiang,China(中国浙江温州) 2020.12 Cultured
Sargassum fusiforme SF25 Nan'ao,Guangdong,China(中国广东南澳) 2020.12 Cultured
Sargassum fusiforme SF26 Wenzhou,Zhejiang,China(中国浙江温州) 2020.12 Wild
Sargassum fusiforme SF27 Wenzhou,Zhejiang,China(中国浙江温州) 2020.12 Cultured
S. pallidum SPA01 Yantai,Shandong,China(中国山东烟台) 2022.06 Wild
S. pallidum SPA02 Yantai,Shandong,China(中国山东烟台) 2022.06 Wild
S. thunbergii ST01 Wenzhou,Zhejiang,China(中国浙江温州) 2021.01 Wild
S. thunbergii ST02 Wenzhou,Zhejiang,China(中国浙江温州) 2020.01 Wild
S. thunbergii ST03 Wenzhou,Zhejiang,China(中国浙江温州) 2020.02 Wild
S. thunbergii ST04 Wenzhou,Zhejiang,China(中国浙江温州) 2020.03 Wild
S. thunbergii ST05 Wenzhou,Zhejiang,China(中国浙江温州) 2020.04 Wild
S. thunbergii ST06 Wenzhou,Zhejiang,China(中国浙江温州) 2020.06 Wild
S. thunbergii ST07 Wenzhou,Zhejiang,China(中国浙江温州) 2022.05 Wild
S. thunbergii ST08 Qingdao,Shandong,China(中国山东青岛) 2022.05 Wild
S. horneri SH01 Wenzhou,Zhejiang,China(中国浙江温州) 2020.02 Wild
S. horneri SH02 Wenzhou,Zhejiang,China(中国浙江温州) 2020.03 Wild
S. horneri SH03 Wenzhou,Zhejiang,China(中国浙江温州) 2020.04 Wild
S. horneri SH04 Wenzhou,Zhejiang,China(中国浙江温州) 2020.05 Wild
S. horneri SH05 Wenzhou,Zhejiang,China(中国浙江温州) 2020.06 Wild
S. horneri SH06 Wenzhou,Zhejiang,China(中国浙江温州) 2022.05 Wild
S. miyabei SM01 Yantai,Shandong,China(中国山东烟台) 2022.05 Wild
S. oligocystum SO01 Trat,Thailand(泰国达叻) 2019.02 Wild
S. oligocystum SO02 Trat,Thailand(泰国达叻) 2019.02 Wild
S. polycystum SP01 Chonburi,Thailand(泰国春武里) 2019.02 Wild
S. ilicifolium SI01 Ranong,Thailand(泰国拉农) 2019.01 Wild
S. binderi SB01 Chantaburi,Thailand(泰国尖竹汶) 2019.01 Wild
Sargassum slices YP01 Shandong,China(中国山东) 2018.05 S. fusiforme
Sargassum slices YP02 Shandong,China(中国山东) 2019.05 S. fusiforme
Sargassum slices YP03 Shandong,China(中国山东) 2021.05 S. fusiforme
Sargassum slices YP04 Guangxi,China(中国广西) 2019.05 S. fusiforme
Sargassum slices YP05 Shandong,China(中国山东) 2015.05 S. crassifolium
Sargassum slices YP06 Zhejiang,China(中国浙江) 2019.11 S. thunbergii
Sargassum slices YP07 Shandong,China(中国山东) 2019.12 S. crassifolium
Sargassum slices YP08 Fujian,China(中国福建) 2019.05 S. miyabei
Sargassum slices YP09 Fujian,China(中国福建) 2017.03 S. sp.
Sargassum slices YP10 Zhejiang,China(中国浙江) 2019.05 S. fusiforme
Sargassum slices YP11 Zhejiang,China(中国浙江) 2019.05 S. fusiforme
Sargassum slices YP12 Zhejiang,China(中国浙江) 2018.05 S. fusiforme
Sargassum slices YP13 Zhejiang,China(中国浙江) 2020.05 S. thunbergii
Sargassum slices YP14 Shandong,China(中国山东) 2015.05 S. horneri
Sargassum slices YP15 Shandong,China(中国山东) 2015.11 S. horneri
), ArticleFig(id=1200147850882544279, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1200147842271637877, language=EN, label=Tab.2, caption=

The verified results on OPLS-DA model of sargassum slices

, figureFileSmall=null, figureFileBig=null, tableContent=
Sample Discriminant index
S. fusiforme Other sargassos
YP01 1.010 1 -0.010 1
YP02 1.097 8 -0.097 8
YP03 1.145 9 -0.145 9
YP04 0.883 0 0.117 0
YP05 -0.232 5 1.232 5
YP06 -0.275 0 1.275 0
YP07 -0.090 3 1.090 3
YP08 -0.392 7 1.392 8
YP09 0.340 1 0.659 9
YP10 1.026 9 -0.026 9
YP11 0.708 3 0.291 7
YP12 0.706 8 0.293 2
YP13 -0.468 2 1.468 2
YP14 -0.070 8 1.070 8
YP15 -0.090 3 1.090 3
), ArticleFig(id=1200147850953847448, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1200147842271637877, language=CN, label=表2, caption=

海藻饮片OPLS-DA模型检验结果

, figureFileSmall=null, figureFileBig=null, tableContent=
Sample Discriminant index
S. fusiforme Other sargassos
YP01 1.010 1 -0.010 1
YP02 1.097 8 -0.097 8
YP03 1.145 9 -0.145 9
YP04 0.883 0 0.117 0
YP05 -0.232 5 1.232 5
YP06 -0.275 0 1.275 0
YP07 -0.090 3 1.090 3
YP08 -0.392 7 1.392 8
YP09 0.340 1 0.659 9
YP10 1.026 9 -0.026 9
YP11 0.708 3 0.291 7
YP12 0.706 8 0.293 2
YP13 -0.468 2 1.468 2
YP14 -0.070 8 1.070 8
YP15 -0.090 3 1.090 3
), ArticleFig(id=1200147851096453789, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1200147842271637877, language=EN, label=Tab.3, caption=

Monounsaturated fatty acid ratio of sargassum slices

, figureFileSmall=null, figureFileBig=null, tableContent=
Sample Fatty acid ratio
C18∶1 n-9/C16∶0 C16∶1 n-7/C16∶0 C16∶1 n-7/C18∶1 n-9 C20∶1 n-9/C16∶1 n-7
S. fusiforme(n=27) 0.20-0.61 0.07-0.30 0.25-0.67 0.43-1.45
YP01 0.24 0.09 0.40 0.96
YP02 0.27 0.09 0.33 1.26
YP03 0.25 0.09 0.36 1.01
YP04 0.33 0.12 0.37 0.71
YP05 0.23 0.07 0.32 0.12
YP06 0.25 0.07 0.29 0.13
YP07 0.50 0.12 0.24 0.07
YP08 0.29 0.12 0.41 0.09
YP09 0.17 0.07 0.40 0.85
YP10 0.27 0.09 0.35 1.17
YP11 0.30 0.13 0.42 0.80
YP12 0.30 0.12 0.40 0.82
YP13 0.24 0.24 0.98 0.20
YP14 0.57 0.28 0.49 0.35
YP15 0.61 0.29 0.49 0.32
), ArticleFig(id=1200147851239060129, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1200147842271637877, language=CN, label=表3, caption=

海藻饮片的单不饱和脂肪酸相关比值

, figureFileSmall=null, figureFileBig=null, tableContent=
Sample Fatty acid ratio
C18∶1 n-9/C16∶0 C16∶1 n-7/C16∶0 C16∶1 n-7/C18∶1 n-9 C20∶1 n-9/C16∶1 n-7
S. fusiforme(n=27) 0.20-0.61 0.07-0.30 0.25-0.67 0.43-1.45
YP01 0.24 0.09 0.40 0.96
YP02 0.27 0.09 0.33 1.26
YP03 0.25 0.09 0.36 1.01
YP04 0.33 0.12 0.37 0.71
YP05 0.23 0.07 0.32 0.12
YP06 0.25 0.07 0.29 0.13
YP07 0.50 0.12 0.24 0.07
YP08 0.29 0.12 0.41 0.09
YP09 0.17 0.07 0.40 0.85
YP10 0.27 0.09 0.35 1.17
YP11 0.30 0.13 0.42 0.80
YP12 0.30 0.12 0.40 0.82
YP13 0.24 0.24 0.98 0.20
YP14 0.57 0.28 0.49 0.35
YP15 0.61 0.29 0.49 0.32
), ArticleFig(id=1200147851478135461, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1200147842271637877, language=EN, label=Tab.4, caption=

Verification of the ratio of C20∶1n-9/C16∶1n-7 in sargassos

, figureFileSmall=null, figureFileBig=null, tableContent=
Species C20∶1
n-9		 C16∶1
n-7		 C20∶1 n-9/
C16∶1 n-7		 Reference
S. fusiforme 1.89 3.71 0.51 [30]
S. fusiforme 2.00 4.49 0.45 [30]
S. fusiforme 2.22 3.03 0.73 [30]
S. fusiforme 2.03 3.01 0.67 [30]
S. fusiforme 2.47 4.04 0.61 [30]
S. fusiforme 2.09 4.93 0.43 [30]
S. ilicifolium 0.20 8.20 0.02 [31]
S. polycystum 0.12 3.81 0.03 [32]
S. fulvellum 0.16 4.03 0.04 [33]
S. horneri 0.08 2.97 0.03 [33]
S. boreale 0.21 4.70 0.04 [33]
S. thunbergii 0.17 5.48 0.03 [33]
S. yezoense 0.10 1.85 0.05 [33]
S. muticum 0.58 6.08 0.10 [34]
S. polyschides 0.07 2.17 0.03 [34]
S. tenerrimum 0.67 4.83 0.14 [35]
S. sp. 0.36 2.74 0.13 [36]
), ArticleFig(id=1200147851608158888, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1200147842271637877, language=CN, label=表4, caption=

文献报道的马尾藻C20∶1 n-9/C16∶1 n-7比值

, figureFileSmall=null, figureFileBig=null, tableContent=
Species C20∶1
n-9		 C16∶1
n-7		 C20∶1 n-9/
C16∶1 n-7		 Reference
S. fusiforme 1.89 3.71 0.51 [30]
S. fusiforme 2.00 4.49 0.45 [30]
S. fusiforme 2.22 3.03 0.73 [30]
S. fusiforme 2.03 3.01 0.67 [30]
S. fusiforme 2.47 4.04 0.61 [30]
S. fusiforme 2.09 4.93 0.43 [30]
S. ilicifolium 0.20 8.20 0.02 [31]
S. polycystum 0.12 3.81 0.03 [32]
S. fulvellum 0.16 4.03 0.04 [33]
S. horneri 0.08 2.97 0.03 [33]
S. boreale 0.21 4.70 0.04 [33]
S. thunbergii 0.17 5.48 0.03 [33]
S. yezoense 0.10 1.85 0.05 [33]
S. muticum 0.58 6.08 0.10 [34]
S. polyschides 0.07 2.17 0.03 [34]
S. tenerrimum 0.67 4.83 0.14 [35]
S. sp. 0.36 2.74 0.13 [36]
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基于特征脂肪酸的中药海藻鉴别研究
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耿玺承 1 , 马善鹏 1, 2 , 陈嘉鹏 1 , 陈璐晶 1 , 林立东 3, * , 刘红兵 1, 2, 4, *
中国药学杂志 | 论著 2024,59(10): 887-895
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中国药学杂志 | 论著 2024, 59(10): 887-895
基于特征脂肪酸的中药海藻鉴别研究
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耿玺承1, 马善鹏1, 2, 陈嘉鹏1, 陈璐晶1, 林立东3, *, 刘红兵1, 2, 4, *
作者信息
  • 1 中国海洋大学, 海洋药物教育部重点实验室, 山东 青岛 266003
  • 2 海洋药物与生物制品功能实验室, 青岛海洋科学与技术试点国家实验室, 山东 青岛 266237
  • 3 温州市洞头区海洋与渔业发展研究中心, 浙江 温州 325700
  • 4 国家药品监督管理局海洋中药质量研究与评价重点实验室, 山东 青岛 266000

    • 耿玺承,女,硕士研究生 研究方向:海洋中药

通讯作者:

*林立东,男,博士,高级工程师 研究方向:基础生物学 Tel: (0577)59386761;
刘红兵,女,博士,教授,博士生导师 研究方向:海洋中药药效物质及质量评价研究 Tel: (0532)82031823
Chemical Identification of TCM Sargassum Based on Characteristic Fatty Acids
Xicheng GENG1, Shanpeng MA1, 2, Jiapeng CHEN1, Lujing CHEN1, Lidong LIN3, *, Hongbing LIU1, 2, 4, *
Affiliations
  • 1 Key Laboratory of Marine Drugs, Ministry of Education, Ocean University of China, Qingdao 266003, China
  • 2 Laboratory for Marine Drugs and Bioproducts, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266237, China
  • 3 Marine and Fishery Development Research Center of Dongtou District, Wenzhou 325700, China
  • 4 NMPA Key Laboratory for Quality Research and Evaluation of Marine Traditional Chinese Medicine, Qingdao 266000, China
出版时间: 2024-05-22 doi: 10.11669/cpj.2024.10.005
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摘要
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目的 利用植物脂质组学和化学分类学开展羊栖菜与其同属易混马尾藻的鉴别研究。方法 收集27批羊栖菜,靶向建立了脂肪酸气相色谱-质谱(GC-MS)指纹图谱;运用主成分分析、正交偏最小二乘判别分析等化学模式识别方法,比较了羊栖菜与同属其他8种22批马尾藻,建立了海藻饮片判别模型;基于羊栖菜与其他马尾藻的差异脂肪酸,结合脂肪酸生合成途径分析,筛选质量标志物。结果 指纹图谱共标定29个共有脂肪酸,相似度均大于0.96。化学模式识别法能够很好区分羊栖菜与同属其他8种22批马尾藻,以此建立的判别模型能够准确验证15批市售海藻饮片。结合差异脂肪酸与脂肪酸生合成途径分析,发现单不饱和脂肪酸在化学分类中的重要作用,提出比值C20∶1 n-9/C16∶1 n-7可作为羊栖菜鉴别的候选质量标志物,14批市售海藻饮片可被准确验证。结论 本研究建立了羊栖菜与同属易混马尾藻的判别模型,发现了脂肪酸质量标志物,为中药海藻真伪鉴别方法的制定提供参考。

中药海藻  /  羊栖菜  /  脂肪酸  /  化学分类学  /  质量标志物

OBJECTIVE To study the identification of Sargassum fusiforme and other sargassos (algae from the genus Sargassum) by targeted lipidomics combined with chemotaxonomy methods. METHODS Twenty-seven batches of S. fusiforme were collected and the GC-MS fingerprint of fatty acids (FAs) was established. The principal component analysis and orthogonal partial least squares discriminant analysis were used to compare S. fusiforme with other 8 species, total 22 batches of sargassos. Based on the differential FAs between S. fusiforme and other sargassos, combined with the analysis of FA biosynthesis pathway, the Q-marker was screened. RESULTS A total of 29 common FAs were identified by the fingerprint, and the similarity was greater than 0.96. The chemical pattern recognition method can well distinguish S. fusiforme from other 8 species, and the discrimination model established by this method can accurately verify 15 batches of commercial slices. Based on the analysis of differential FAs and FA biosynthesis pathway, monounsaturated fatty acid was found to play an important role in chemical classification. The ratio C20∶1 n-9/C16∶1 n-7 was proposed to be used as a candidate Q-marker for the identification of S. fusiforme and 14 batches of commercial slices can be accurately verified. CONCLUSION This study establishes the identification model of S. fusiforme and other confused sargassos, finds the Q-marker of FAs, and provides a reference for the establishment of the identification method of TCM SARGASSUM.

TCM sargassum  /  Sargassum fusiforme  /  fatty acid  /  chemotaxonomy  /  Q-marker
耿玺承, 马善鹏, 陈嘉鹏, 陈璐晶, 林立东, 刘红兵. 基于特征脂肪酸的中药海藻鉴别研究. 中国药学杂志, 2024 , 59 (10) : 887 -895 . DOI: 10.11669/cpj.2024.10.005
Xicheng GENG, Shanpeng MA, Jiapeng CHEN, Lujing CHEN, Lidong LIN, Hongbing LIU. Chemical Identification of TCM Sargassum Based on Characteristic Fatty Acids[J]. Chinese Pharmaceutical Journal, 2024 , 59 (10) : 887 -895 . DOI: 10.11669/cpj.2024.10.005
正文
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中药海藻(sargassum)始载于《神农本草经》,性寒、味咸,功能软坚散结,消痰利水,常用于治疗高血脂、冠心病、肿瘤、乳腺增生、淋巴结核等[1-4],近年来受到国内外学者较多关注[5-9]。据2020年版《中国药典》一部,中药海藻为褐藻门马尾藻属(Sargassum)海蒿子[S. pallidum (Turn.) C. Ag]或羊栖菜[S. fusiforme (Harv.) Setch]的干燥藻体。其中,羊栖菜是历史习用品种[2-3],近年来养殖产量持续位居我国海水藻类第五位[10],已成为中药海藻的主流药用品种。
马尾藻属(Sargassum)在我国沿海物种多样性丰富[11-12],但这也导致海藻药材品种较为混乱,常见伪品有同属的鼠尾藻、铜藻、海黍子等[13-15]。马尾藻属物种鉴定一直是藻类分类学研究的难点,因藻体形态和器官组织结构相近、叶和气囊表型随环境变化,生理特征趋同等特点,使得传统的性状鉴别、显微鉴别等方法很难直接鉴定到该属具体物种。DNA分子标记技术准确可靠,尽管已有关于马尾藻属分类修订的研究[16-17],但距离形成海藻药材DNA分子鉴定体系尚远。鉴于目前海藻真伪鉴别方法有限,课题组聚焦主流品种羊栖菜,从植物脂质组学和化学分类学的角度展开真实性评价相关的理化鉴别研究。
脂质在生物学过程中发挥着广泛而重要的作用。作为代谢组学的分支之一,脂质组学研究在中药药效物质及作用机制研究中已有较多应用[18]。植物脂质组学能够系统解析植物体脂质组成与表达变化,但甚少见有基于植物脂质组学的中药鉴定研究。马尾藻属的脂质主要包括脂肪酸及其衍生脂(磷脂、糖脂、甘油酯等)、甾醇以及类胡萝卜素类等,具有抗氧化、降血脂、抗炎免疫等生物活性[19]。其中,脂肪酸是人体营养和健康所必需成分[20],是重要的活性成分。目前已有脂肪酸作为大型海藻分类标记物的报道[21-23],本研究基于前期工作聚焦脂肪酸[24],靶向建立了羊栖菜气相色谱-质谱(GC-MS)指纹图谱,通过化学模式识别比较羊栖菜与同属其他马尾藻的差异,基于脂肪酸生合成途径探讨可用于鉴别羊栖菜的候选质量标志物,以期为中药海藻真伪鉴别提供简单、准确、快速的方法。
1 仪器与材料
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1.1 仪器
Thermo Trace 1300型气相色谱-ISQ 型单四级杆质谱联用系统(美国Thermo Fisher公司);Omegawax 250毛细管色谱柱(0.25 mm×30 m,0.25 μm,美国Supelco公司);SOX-606型脂肪测定仪(山东海能科学仪器有限公司);MS3 basic型涡旋振荡器(广东 IKA公司);BSM-2204型电子天平(上海卓精电子科技有限公司)。
1.2 试剂
37种脂肪酸甲酯混标(纯度>99%,批号XA22213V,美国Sigma-Aldrich公司);天然鱼油(澳大利亚Swisse公司,每1 500 mg天然鱼油中含二十二碳六烯酸180 mg和二十二碳五烯酸270 mg)。正己烷为色谱纯;二氯甲烷、甲醇、浓盐酸、氢氧化钾为分析纯。
1.3 药材
共收集羊栖菜27批,同属其他8种马尾藻共22批。其中,羊栖菜(S. fusiforme)、鼠尾藻(S. thunbergii)以及铜藻(S. horneri)(温州市洞头区海洋与渔业发展研究中心林立东高工提供并鉴定);海蒿子(S. pallidum)和海黍子(S. miyabei)(中国科学院海洋所孙忠民博士提供并鉴定);少囊马尾藻(S. oligocystum)、匍枝马尾藻(S. polycystum)、冬青叶马尾藻(S. ilicifolium)、宾德马尾藻(S. binderi)(泰国农业大学Mr. Nattasak Suwannasingha和Dr. Jantana Praiboon提供并鉴定)。海藻饮片15批(市售),经中国海洋大学刘红兵教授鉴定,具体信息见表1。
2 方法与结果
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2.1 溶液的制备
供试品溶液的制备:取海藻粉末约0.75 g,精密称定后,置于脂肪测定仪,加入二氯甲烷-甲醇(2∶1)120 mL,90 ℃提取10次;提取液减压浓缩,干燥得总脂。取总脂,加入1 mol·L-1的KOH甲醇溶液1 mL,70 ℃热皂化10 min;再加入1 mol·L-1盐酸甲醇溶液2 mL,甲酯化衍生10 min。在反应液中加入400 μL去离子水和600 μL正己烷,混匀,吸取有机层;共萃取3次,合并有机层,即得供试品溶液。4 ℃保存,待GC-MS分析。
对照品溶液的制备:取25 μL鱼油,按供试品溶液制备方法进行皂化和衍生化后,取10 μL,加入10 μL 37种脂肪酸甲酯混标和20 μL正己烷,混匀,即得脂肪酸甲酯对照品溶液。
2.2 GC-MS条件
使用Omegawax 250毛细管色谱柱(0.25 mm×30 m,0.25 μm,美国Supelco公司)。进样口温度250 ℃,炉温180 ℃;采用程序升温,初始温度设定180 ℃并保持3 min,再以2.5 ℃·min-1升至240 ℃,保持2 min。载气为氦气,流速为1 mL·min-1;采用分流模式,分流比为20∶1;进样体积1 μL。质谱离子源为EI源,温度280 ℃,能量70 eV,全扫描模式,范围m/z 35~550。使用Xcalibur2.0处理原始数据,采用面积归一化法计算色谱峰相对含量。
2.3 羊栖菜脂肪酸指纹图谱
2.3.1 方法学考察
精密度实验:精密称取羊栖菜SF13粉末,按“2.1”项下方法制备供试品溶液,按“2.2”项下条件连续进样6次,以6号峰(棕榈酸)为参照,计算共有峰的相对保留时间相对标准偏差(RSD)均小于1.0%,相对峰面积RSD均小于5.0%,表明仪器精密度良好。
重复性实验:精密称取羊栖菜SF13粉末,按“2.1”项下方法平行制备6份供试品溶液,按“2.2”项下条件测定,以6号峰(棕榈酸)为参照,计算共有峰的相对保留时间RSD均小于1.0%,相对峰面积RSD均小于5.0%,表明方法重复性良好。
稳定性实验:精密称取羊栖菜SF13粉末,分别于制备供试品溶液后0、2、4、6、8、12、24 h时,按“2.2”项下条件测定,以6号峰(棕榈酸)为参照,计算共有峰的相对保留时间RSD均小于1.0%,相对峰面积RSD均小于5.0%,表明供试品溶液在24 h内稳定性良好。
2.3.2 指纹图谱建立
本研究收集的27批羊栖菜样品包括野生品种及养殖品种(又分为3个品系);采收时间从12月至第二年5月收菜,涵盖了羊栖菜藻体的主要生长时期;产地分布从南至北,涉及广东、浙江、山东等3省主要产区,具有较强的代表性。取上述27批羊栖菜,按“2.1”项下方法制备供试品溶液,按“2.2”项下条件测定。将数据导入《中药色谱指纹图谱相似度评价系统》(2012版),选择中位数矢量法、时间窗宽度为0.1 min,经多点校正后进行峰匹配,生成对照指纹图谱,标定了29个脂肪酸共有峰,并通过与对照品比对进行鉴定(图1)。其中,棕榈酸(C16∶0,峰6)保留时间居中、分离度良好,选定为参比峰。计算27批羊栖菜的指纹图谱与对照图谱的相似度,相似度大于0.96,符合一致性要求。指纹图谱相似度结果提示,羊栖菜脂肪酸组成较为稳定。
2.4 化学模式识别比较
2.4.1 主成分分析
取22批同属他种马尾藻,按“2.1”项下方法制备供试品溶液,按“2.2”项下条件测定。以29个脂肪酸共有峰的相对含量为变量,采用SIMCA 14.1对27批羊栖菜和22批同属马尾藻进行无监督模式的主成分分析(principal component analysis,PCA),前六个主成分累计方差贡献率79.6%,能够解释大部分样品信息的变化。从PCA得分图可见(图2),羊栖菜与大多数同属他种马尾藻可以明显分区;不能与羊栖菜区分的4个马尾藻(ST01,ST02,ST04和ST05)均为鼠尾藻。
2.4.2 正交偏最小二乘判别分析
为进一步对羊栖菜与同属他种马尾藻进行有效区分,运用SIMCA 14.1进行有监督的正交偏最小二乘判别分析(orthogonal partial least squares discriminant analysis,OPLS-DA),R2X=82.7%,R2Y=92.7%,Q2=87.5%,说明模型具有较好的解释及预测能力。200次置换检验结果说明模型未过拟合(图3A)。OPLS-DA结果表明,羊栖菜及同属他种马尾藻明显分为两类(图3B),提示两者脂肪酸组成存在显著差异。以变量投影重要性(variable importance projection, VIP)值>1.0为标准筛选差异脂肪酸(图3C),得到C16∶0、C20∶5 n-3、C18∶3 n-3、C18∶4 n-3、C16∶1 n-7、C18∶1 n-9、C18∶2 n-6、C20∶4 n-6、C20∶3 n-6等9个差异脂肪酸,具体见图3D。
使用15批海藻饮片验证建立的OPLS-DA模型,以0.5作为判别指数阈值,所有判别结果正确(表2)。其中,YP09实际为几种马尾藻混杂的饮片样品,其判别指数与羊栖菜及同属他种马尾藻的判别指数比较接近,结果与实际情况较为相符。
2.5 分类标志物探讨
脂肪酸是生物膜的重要组成部分,其生合成由遗传物质调控,具有作为化学分类标志物的潜质。
脂肪酸合成从乙酰辅酶A开始,见图4,在酶的催化下经碳循环延长,形成含有16和18个碳的饱和脂肪酸(saturated fatty acid, SFA),即棕榈酸(C16∶0)和硬脂酸(C18∶0);继而在Δ9脂肪酸去饱和酶的作用下,分别合成单不饱和脂肪酸(monounsaturated fatty acid, MUFA),如棕榈油酸(C16∶1 n-7)或油酸(C18∶1 n-9);棕榈油酸可以进一步介导n-7系列MUFA的合成;油酸主要有两条途径,或是参与n-9系列MUFA的合成例如生成花生烯酸(C20∶1 n-9),或是参与多不饱和脂肪酸(polyunsaturated fatty acid, PUFA)的生成,包括n-6 PUFA(亚油酸等)和n-3 PUFA(亚麻酸)等[25-29]。
从上述分析可知,MUFA在脂肪酸生合成途径中扮演着重要的承上启下角色。例如,棕榈油酸能够反映n-7 MUFA的合成水平,油酸能直接及间接反映n-9 MUFA以及PUFA的合成水平。鉴于上述2个MUFA分子同时是羊栖菜与同属他种马尾藻的关键差异脂肪酸,故结合脂肪酸生合成途径,基于脂肪酸比值寻找分类标志物。比值C18∶1 n-9/C16∶0、C16∶1 n-7/C16∶0、C16∶1 n-7/C18∶1 n-9和C20∶1 n-9/C16∶1 n-7在羊栖菜与鼠尾藻/铜藻中存在显著差异(P<0.01)。从具体数值来看,比值C20∶1 n-9/C16∶1 n-7能够明显区分本研究中的羊栖菜与同属他种马尾藻(图5),而且能够正确判别饮片YP09以外的其他14批饮片(表3)。YP09是含有羊栖菜的掺伪品,可以利用建立的OPLS-DA模型准确判别。
此外,本研究还采用文献报道的马尾藻属脂肪酸数据[30-36],对提出的C20∶1 n-9/C16∶1 n-7比值进行验证。从表4可以看出,该比值能够区分羊栖菜与同属马尾藻。以上结果提示,C20∶1 n-9/C16∶1 n-7比值可作为羊栖菜鉴别的候选标志物。需要注意的是,马尾藻中常含有C20∶1 n-9的同分异构体C20∶1 n-11,文献[37-38]采用的脂肪酸测定方法不能将两者完全分离,导致C20∶1 n-9/C16∶1 n-7比值偏差很大。
从高等植物到藻类,都有利用脂肪酸比值进行化学分类研究的报道,例如Barthetca发现n-7/n-9 MUFA可用于区分芸苔属植物[39],Wu等[40]利用C18∶1 n-9/C18∶2 n-6判别掺假油料作物,Khotimchenko等[41-42]报道18∶1 n-7/18∶1 n-9、C20∶5 n-3/C20∶4 n-6分别可用于绿藻门、红藻门的分类研究。
3 结论与讨论
收起
中药海藻是2020年版《中国药典》一部收载的2种海洋来源藻类中药材之一(另一为昆布)。海蒿子是中药海藻的基原植物之一,主要分布于我国黄渤海近岸岛屿与岛礁,因野生资源量逐年减少,现市场很少见到该种药材。羊栖菜在我国已形成规模化养殖[43],是当前中药海藻的主流品种,但常有同属他种马尾藻冒充,缺乏简单有效的专属性鉴别方法。
马尾藻属藻类,常统称Sargassos,属于褐藻门(Phaeophyta)墨角藻目(Fucales)马尾藻科(Sargassaceae),成熟藻体由假根、茎(主茎、侧生茎)、叶 (气囊)和生殖托(雄托、雌托)等组成。藻体形态受生长周期、环境因素等影响巨大,致使自1820年建属以来,物种鉴别一直是研究难点。完整藻体尚难以区分物种,切段制成饮片后,其鉴别难度更甚,严重影响了中药海藻临床用药的有效性和安全性。本研究以羊栖菜为研究对象,建立了靶向脂肪酸GC-MS指纹图谱,可通过OPLS-DA准确区分羊栖菜与同属他种马尾藻;基于马尾藻物种间的差异脂肪酸,结合生合成分析,首次提出比值C20∶1 n-9/C16∶1 n-7可作为羊栖菜鉴别的候选质量标志物。本研究尚存在样品数量有限等诸多不足之处,结果也有待于大批量样本的检验。研究旨在抛砖引玉,希望引起广大同行对海洋中药质量的关注,共同建设我国蓝色药库。
基金
收起
  • 国家自然科学基金项目资助(81973433)
  • 国家药品监督管理局海洋中药质量研究与评价重点实验室课题资助(HYZY-ZZKT-2022002)
  • 青岛海洋科技中心专项资助(2022QNLM030003-2)
文献
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2024年第59卷第10期
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doi: 10.11669/cpj.2024.10.005
  • 接收时间：2023-03-29
  • 首发时间：2025-11-25
  • 出版时间：2024-05-22
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  • 收稿日期：2023-03-29
基金
国家自然科学基金项目资助(81973433)
国家药品监督管理局海洋中药质量研究与评价重点实验室课题资助(HYZY-ZZKT-2022002)
青岛海洋科技中心专项资助(2022QNLM030003-2)
作者信息
    1 中国海洋大学, 海洋药物教育部重点实验室, 山东 青岛 266003
    2 海洋药物与生物制品功能实验室, 青岛海洋科学与技术试点国家实验室, 山东 青岛 266237
    3 温州市洞头区海洋与渔业发展研究中心, 浙江 温州 325700
    4 国家药品监督管理局海洋中药质量研究与评价重点实验室, 山东 青岛 266000

通讯作者:

*林立东,男,博士,高级工程师 研究方向:基础生物学 Tel: (0577)59386761;
刘红兵,女,博士,教授,博士生导师 研究方向:海洋中药药效物质及质量评价研究 Tel: (0532)82031823
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2种不同金属材料的力学参数
科
Family		 属数
Number of
genus		 种数
Number of
species		 占总种数比例
Percentage of
total species (%)		 属
Genus		 种数
Number of
species		 占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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